佛波酯对CKI p15^(INK4B)高表达的人黑色素瘤细胞生长及凋亡的影响  

Effect of PMA on Growth and Apoptosis of Human Melanoma Cell Overexpressing CKI p15^(INK4B)

作　　者：童迎凯[1] 柳惠图[1] 刘军[1] 

机构地区：[1]北京师范大学生命科学学院

出　　处：《生物化学与生物物理进展》2001年第2期198-202,共5页Progress In Biochemistry and Biophysics

基　　金：国家自然科学基金!资助项目 (39780 0 14 ) ;北京市自然科学基金!重点项目 (796 10 0 1) ;国家重点基础研究项目!(19990 5 390 1) &&

摘　　要：通过DNA体外重组和转染技术 ,将已构建好的含有p15基因全长的质粒 pXJ 41 p15转染p15缺失的人黑色素瘤细胞A375 ,经G418筛选出阳性单克隆 ,并经PCR、蛋白质印迹等检测 ,证明建立了 p15稳定高表达的细胞模型 .实验表明 ,经佛波酯 (PMA)长期处理 ,实验组细胞中的 p15表达水平进一步上升 ,同时观察到蛋白激酶C (PKC)活性下降 ,与对照组细胞相比 p15高表达的细胞PKC活性及细胞生长均受到较强烈的抑制 ,并能引起约 30 %的实验组细胞发生凋亡 .凋亡细胞中Caspase3P2 0亚基的水平上升 .结果表明 ,CKIp15与PKC信号系统在调节细胞增殖、凋亡的过程中具有一定的相关性 ,它们的协同作用可能启动了细胞中与Caspase相关的凋亡通路 ,使细胞发生凋亡 .The plasmid pXT-41-p15, which contains the full length DNA coding for p15 was introduced into human melanoma cell line A375 in which p15 was deleted by DNA recombination and transfection. Using G418, the positive clones were selected. And the cell model overexpressing p15 was constructed successfully through the analysis of PCR and Western blot. It is showed that the expression of p15 was further enhanced after the cells overexpressing p15 were treated with PMA for 72 hours. In contrast of the control cells, the PKC activity was further declined in the cells overexpressing p15 after treated with PMA. At the same time, the growth rate of cell was decreased more significantly and approximate 30% apoptotic cells were found. The expression of Caspase-3 (P20) was increased in the apoptotic cells. It is indicated that CKI p15 was related to PKC signal transduction in the regulation of cell proliferation and apoptosis. They may be involved in the apoptotic pathway including Caspase3, thus inducing the apoptosis of cells.

关 键 词：P15 周期蛋白依赖激海抑制因子 佛波酯 蛋白激酶C 细胞凋亡 

分 类 号：Q253[生物学—细胞生物学] Q26